Primorigen Biosciences SBIR Proposal Abstract Primorigen Biosciences will develop a defined, xenobiotic-free synthetic cell culture matrix that fully supports maintenance and directed differentiation of human induced pluripotent stem cells (hiPSCs) down all three germ layers and dramatically reduces the costs of stem cell culture processes. The current, most commonly used matrices either create significant reliability and safety issues, or are effective only with certain cell lines given their limited mechanisms for cel adhesion. To address these drawbacks, Primorigen will develop a new synthetic cell culture matrix that recapitulates key attributes of natural extracellular matrix by presenting diverse synthetic ligands that offer numerous mechanisms for cell adhesion and facilitate sequestration of endogenous factors thereby reducing the need for and costs of using exogenous growth factors. Phase I studies will demonstrate feasibility by (a) creating a library of synthetic cell adhesion and endogenous factor sequestering ligands, (b) optimizing the coating conditions of ligand mixtures onto polystyrene surfaces, and (c) validating the new synthetic cell culture matrix for hiPSC maintenance and directed differentiation into various lineages. Phase II objectives will build on this work by expanding the synthetic ligand mixture approach to validate their use in large scale production and differentiation of hiPSCs. Commercial outputs will include (i) a broadly applicable synthetic Matrigel replacement comprised of synthetic modular ligand mixtures optimized for stem cell maintenance and differentiation, (ii) polystyrene tissue culture platforms (e.g., Petri dish, multiwell plate and microcarrier) pre-coated with the synthetic modular ligand mixtures, and (iii) a library of synthetic modular ligands from which made to order, customized mixtures can be assembled according to individual customers' unique needs.